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Journal of Virology, December 2007, p. 12872-12880, Vol. 81, No. 23
0022-538X/07/$08.00+0     doi:10.1128/JVI.00974-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Virp1 Is a Host Protein with a Major Role in Potato Spindle Tuber Viroid Infection in Nicotiana Plants{triangledown} ,{dagger}

K. Kalantidis,1* M. A. Denti,1,{ddagger} S. Tzortzakaki,1 E. Marinou,2 M. Tabler,1,§ and M. Tsagris2*

Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology-Hellas, P.O. Box 1527, GR-71110 Heraklion, Crete, Greece,1 Department of Biology, University of Crete, Heraklion, Crete, Greece2

Received 6 May 2007/ Accepted 16 September 2007

Viroids are small, circular, single-stranded RNA molecules that, while not coding for any protein, cause several plant diseases. Viroids rely for their infectious cycle on host proteins, most of which are likely to be involved in endogenous RNA-mediated phenomena. Therefore, characterization of host factors interacting with the viroid may contribute to the elucidation of RNA-related pathways of the hosts. Potato spindle tuber viroid (PSTVd) infects several members of the Solanaceae family. In an RNA ligand screening we have previously isolated the tomato protein Virp1 by its ability to specifically interact with PSTVd positive-strand RNA. Virp1 is a bromodomain-containing protein with an atypical RNA binding domain and a nuclear localization signal. Here we investigate the role of Virp1 in the viroid infection cycle by the use of transgenic lines of Nicotiana tabacum and Nicotiana benthamiana that either overexpress the tomato Virp1 RNA or suppress the orthologous Nicotiana genes through RNA silencing. Plants of the Virp1-suppressed lines were not infected by PSTVd or Citrus exocortis viroid through mechanical inoculation, indicating a major role of Virp1 in viroid infection. On the other hand, overexpression of tomato Virp1 in N. tabacum and N. benthamiana plants did not affect PSTVd KF 440-2 infectivity or symptomatology in these species. Transfection experiments with isolated protoplasts revealed that Virp1-suppressed cells were unable to sustain viroid replication, suggesting that resistance to viroid infection in Virp1-suppressed plants is likely the result of cell-autonomous events.

* Corresponding author. Mailing address for Kriton Kalantidis: IMBB-FORTH, Vasilika Vouton, 71110, P.O. Box 1527, Heraklion, Crete, Greece. Phone: 302810 394364. Fax: 302810 394408. E-mail: kriton{at}imbb.forth.gr. Mailing address for Mina Tsagris: Department of Biology, University of Crete, Vasilika Vouton, P.O. Box 2208, GR-71409, Heraklion, Crete, Greece. Phone: 302810 394367. Fax: 302810 394404. E-mail: tsagris{at}imbb.forth.gr

{triangledown} Published ahead of print on 26 September 2007.

{dagger} Supplemental material for this article may be found at http://jvi.asm.org/.

{ddagger} Present address: Centre for Integrative Biology (CIBIO), University of Trento, via delle Regole, 101, 38060 Mattarello (TN), Italy.

§ Deceased on 3 April 2005.

Journal of Virology, December 2007, p. 12872-12880, Vol. 81, No. 23
0022-538X/07/$08.00+0     doi:10.1128/JVI.00974-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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